The Optimized Delivery of Triterpenes by Liposomal Nanoformulations: Overcoming the Challenges.

The last decade has witnessed a sustained increase in the research development of modern-day chemo-therapeutics, especially for those used for high mortality rate pathologies. However, the therapeutic landscape is continuously changing as a result of the currently existing toxic side effects induced by a substantial range of drug classes. One growing research direction driven to mitigate such inconveniences has converged towards the study of natural molecules for their promising therapeutic potential. Triterpenes are one such class of compounds, intensively investigated for their therapeutic versatility. Although the pharmacological effects reported for several representatives of this class has come as a well-deserved encouragement, the pharmacokinetic profile of these molecules has turned out to be an unwelcomed disappointment. Nevertheless, the light at the end of the tunnel arrived with the development of nanotechnology, more specifically, the use of liposomes as drug delivery systems. Liposomes are easily synthesizable phospholipid-based vesicles, with highly tunable surfaces, that have the ability to transport both hydrophilic and lipophilic structures ensuring superior drug bioavailability at the action site as well as an increased selectivity. This study aims to report the results related to the development of different types of liposomes, used as targeted vectors for the delivery of various triterpenes of high pharmacological interest.

Methotrexate and Cetuximab-Biological Impact on Non-Tumorigenic Models: In Vitro and In Ovo Assessments.

Background Objectives: The neoplastic process remains a major health problem facing humanity. Although there are currently different therapeutic options, they raise a multitude of shortcomings related to the toxic effects associated with their administration. Methotrexate (Met) and Cetuximab (Cet) are two basic chemotherapeutics used in cancer practice, but notwithstanding despite many years of use, the mechanisms by which the multitude of side-effects occur are not yet fully understood. Thus, the present study focused on the in vitro and in ovo evaluation of the associated toxic mechanisms on keratinocytes, keys cells in the wound healing process. Materials and Methods: The two chemotherapeutics were tested in eight different concentrations to evaluate keratinocytes viability, the anti-migratory effect, and the influence on the expression of markers involved in the production of cell apoptosis. In addition, the potential irritating effect on the vascular plexus were highlighted by applying the in ovo method, chick chorioallantoic membrane (HET-CAM). Results: The results revealed that Met induced decreased cell viability as well as increased expression of pro-apoptotic genes. In the vascular plexus of the chorioallantoic membrane, Met caused vascular irritation accompanied by capillary hemorrhage and vascular stasis. Conclusions: Summarizing, Cet presents a safer toxicological profile, compared to Met, based on the results obtained from both in vitro (cell viability, wound healing, RT-PCR assays), and in ovo (HET-CAM assay) techniques.

Impact of Vitamin D on Osseointegration in Dental Implants: A Systematic Review of Human Studies.

This systematic review evaluates the impact of Vitamin D levels on dental implant osseointegration, hypothesizing that optimal Vitamin D enhances success rates, and aims to synthesize data on its relationship with clinical outcomes in implantology. A comprehensive search across PubMed, Cochrane Library, and Web of Science databases included seven peer-reviewed articles meeting the criteria for the review. These studies, conducted between 2008 and 2021, included human subjects and explicitly correlated serum Vitamin D levels with dental implant outcomes, following PRISMA guidelines. The selected studies involved 1462 participants and examined 4450 dental implants. Key findings included a varied implant loss rate ranging from 3.9% to 11.4% across the studies. One study reported a 9.8% implant loss rate, yet found no significant association between Vitamin D receptor polymorphism and implant success. Another study indicated successful implantation following Vitamin D3 supplementation, even in severe deficiency cases. The highest implant loss rate (11.1%) was observed in severely Vitamin D-deficient patients, particularly when compounded by risk factors such as smoking and periodontal disease. Additionally, one study noted significantly improved bone density following post-surgical Vitamin D supplementation for up to 12 weeks. The review supports a link between sufficient Vitamin D levels and successful dental implant osseointegration, suggesting Vitamin D deficiency as a potential risk factor for increased failure and advocating for Vitamin D evaluations in pre-surgical planning to potentially enhance implantology outcomes.

Efficacy and Safety of Botulinum Toxin in the Management of Temporomandibular Symptoms Associated with Sleep Bruxism: A Systematic Review.

Sleep bruxism, characterized by involuntary grinding or clenching of teeth during sleep, poses significant challenges in management due to its potential to induce temporomandibular joint disorders (TMDs) and other related symptoms. The use of Botulinum toxin Type A (BoNT-A), also known as Botox®, has been proposed as a therapeutic intervention. This systematic review aims to evaluate the efficacy and safety of BoNT-A in the management of sleep bruxism, focusing on pain reduction, improvement in jaw function, reduction in bruxism episodes, and the incidence of adverse effects. An exhaustive search was conducted across PubMed, Scopus, and Embase databases up to January 2024, adhering to the PRISMA guidelines. Nine randomized clinical trials (RCTs) involving 137 participants were analyzed for efficacy and safety outcomes. The studies demonstrated a significant reduction in mean pain scores (from 7.1 to 0.2 at 6 months and 1 year post-treatment in one study) and a notable decrease in the number of bruxism events (from 4.97/h to 1.70/h in the BoNT-A group in another study). Additionally, improvements were observed in jaw stiffness and total sleep time. Adverse effects varied but were generally mild and transient, including injection site pain in 20% of participants in one study and cosmetic changes in smile in 15.4% of patients in another. These findings suggest that BoNT-A injections may provide some benefits for treating nocturnal bruxism, potentially reducing TMD symptoms like pain and improving jaw function. However, these findings are preliminary due to variability in study designs and the absence of detailed statistical analysis.

A Systematic Review of the Relationship between Serum Vitamin D Levels and Caries in the Permanent Teeth of Children and Adolescents.

This systematic review critically evaluates the association between serum Vitamin D levels and dental caries incidence in the permanent teeth of children and adolescents. The search strategy comprised three databases (PubMed, Scopus, Embase), up to November 2023, targeting studies on the correlation between Vitamin D and dental caries in permanent dentition. The eligibility criteria focused on observational studies involving children and adolescents aged 12 to 19 years with permanent dentition. The screening process, guided by the PRISMA guidelines and the Newcastle-Ottawa Scale for quality assessment, resulted in the inclusion of eight studies conducted across various global regions from 2013 to 2023. The analysis revealed that Vitamin D insufficiency and deficiency were prevalent among the study populations, ranging from 17.3% to 69.4%. Specifically, children and adolescents with Vitamin D insufficiency (<50 nmol/L) were found to have significantly higher odds of developing caries, with odds ratios (ORs) ranging from 1.13 to 2.57. Conversely, two studies indicated a protective effect of higher Vitamin D levels, with an OR of 0.80 and 0.59, respectively, for caries among children and adolescents with serum levels ≥ 50 nmol/L, suggesting an inverse relationship between Vitamin D status and caries risk. The results indicate both the protective role of adequate serum levels of Vitamin D above 20 ng/mL and the increased risk associated with insufficient levels below this threshold. However, the variations in study quality, methodologies and geographic settings underscore the challenges in drawing universal conclusions. Despite these limitations, our review suggests that improving Vitamin D status could be a beneficial component of preventive strategies against dental caries in children and adolescents, warranting further research to clarify the clinical significance of our findings.

Rutin Linoleate Triggers Oxidative Stress-Mediated Cytoplasmic Vacuolation in Non-Small Cell Lung Cancer Cells.

Lung cancer (LC) represents one of the most prevalent health issues globally and is a leading cause of tumor-related mortality. Despite being one the most attractive compounds of plant origin due to its numerous biological properties, the therapeutic applications of rutin (RUT) are limited by its disadvantageous pharmacokinetics. Thus, the present study aimed to evaluate in vitro the application of two RUT fatty acids bioconjugates, rutin oleate (RUT-O) and rutin linoleate (RUT-L), as potential improved RUT-based chemotherapeutics in non-small cell lung cancer (NSCLC) treatment. The results indicate that both compounds lacked cytotoxic potential in EpiAirway™ tissues at concentrations up to 125 µM. However, only RUT-L exerted anti-tumorigenic activity in NCI-H23 NSCLC cells after 24 h of treatment by reducing cell viability (up to 47%), proliferation, and neutral red uptake, causing cell membrane damage and lactate dehydrogenase (LDH) leakage, affecting cytoskeletal distribution, inducing cytoplasmic vacuolation, and increasing oxidative stress. The cytopathic effects triggered by RUT-L at 100 and 125 µM are indicators of a non-apoptotic cell death pathway that resembles the characteristics of paraptosis. The novel findings of this study stand as a basis for further investigations on the anti-cancer properties of RUT-L and their underlying mechanisms.

Comprehensive in vitro and in ovo assessment of cytotoxicity: Unraveling the impact of sodium fluoride, xylitol, and their synergistic associations in dental products.

Over the past several decades, dental health products containing fluoride have been widely employed to mitigate tooth decay and promote oral hygiene. However, concerns regarding the potential toxicological repercussions of fluoride exposure have incited continuous scientific inquiry. The current study investigated the cytotoxicity of sodium fluoride (NaF) and xylitol (Xyl), both individually and in combination, utilizing human keratinocyte (HaCaT) and osteosarcoma (SAOS-2) cell lines. In HaCaT cells, NaF decreased proliferation in a concentration-dependent manner and induced apoptosis-related morphological changes at low concentrations, whereas Xyl exhibited dose-dependent cytotoxic effects. The combination of NaF and Xyl reduced cell viability, particularly at higher concentrations, accompanied by apoptosis-like morphological alterations. Sub-cytotoxic NaF concentrations (0.2%) significantly affected caspase activity and the expression of pro-apoptotic genes. Conversely, Xyl demonstrated no discernible effect on these biological parameters. In SAOS-2 cells, NaF increased proliferation at high concentrations, contrasting with Xyl's concentration-dependent cytotoxic effects. The combination of NaF and Xyl had a minimal impact on cell viability. Sub-cytotoxic NaF concentrations did not influence caspase activity or gene expression, while Xyl induced dose-dependent morphological alterations, increased caspase activity, and upregulated pro-apoptotic gene expression. In ovo experiments on the chorioallantoic membrane (CAM) revealed that only NaF induced irritant effects, suggesting potential vascular adverse outcomes. This study advocates for the combined use of NaF and Xyl, highlighting their cytotoxicity benefits in healthy cells while maintaining safety considerations for tumor cells.

Examining the Role of Vitamin D in Caries Susceptibility in Children's Deciduous Teeth: A Systematic Review.

The global prevalence of dental caries in deciduous teeth remains a significant health concern, affecting almost 70% of children by the age of six in specific regions. This systematic review aspired to methodically investigate the association between vitamin D levels and susceptibility to caries in children's deciduous teeth. A detailed search, guided by the PRISMA and PROSPERO guidelines, was conducted across three prominent electronic databases: PubMed, Web of Science, and Scopus, culminating in August 2023. The search integrated various keywords related to vitamin D and dental caries in primary dentition, yielding an initial pool of 1678 articles. After meticulous scrutiny, seven studies with a total of 7655 participants were deemed suitable for inclusion. The studies represented diverse geographical regions, showcasing varied vitamin D levels and sun exposure. Patient habits like brushing frequency, dental visits, and vitamin consumption also varied across studies. The analysis pinpointed vitamin D deficiency as a potential risk factor in some of the studies, with Odds Ratios (OR) ranging from 0.68 to 2.15. Statistically significant associations between vitamin D deficiency and caries susceptibility were documented in three studies (ORs of 2.15, 1.98, and 1.70). This comprehensive review elucidates the complex relationship between vitamin D levels and dental caries in children's deciduous teeth. While some studies spotlight vitamin D's pivotal role in dental health, inconsistencies across studies and regional differences necessitate more in-depth, globally representative investigations. Ensuring optimal vitamin D levels may play an integral role in dental health strategies. However, it is important to highlight that the roles of these studied factors might differ in deciduous teeth compared to permanent teeth.

Validation of the Romanian Version of the Halitosis Associated Life-Quality Test (HALT) in a Cross-Sectional Study among Young Adults.

Halitosis is a common condition with a significant impact on individuals' quality of life. The Halitosis Associated Life-Quality Test (HALT) is a reliable instrument for measuring this impact. This study aimed to introduce and validate the Romanian translation of the HALT questionnaire (R-HALT). We hypothesized that the R-HALT would demonstrate good reliability and validity in measuring the impact of halitosis on quality of life among Romanian teenagers and young adults. Our objectives were to translate and adapt the HALT, validate it among a cross-sectional group, and evaluate the extent of halitosis in this population. A multicentric cross-sectional design followed, which was approved by the Ethics Research Committee in Romania. The translation process involved independent translations, retro-translations, expert review, and pre-testing. The psychometric properties were evaluated among 150 patients (mean age 23.6 ± 1.8; 51% males) at dental clinics, including reliability, convergent, and discriminant validity, using accepted statistical measures such as Cronbach's alpha and Intraclass Correlation Coefficient (ICC). The R-HALT revealed strong internal consistency with Cronbach's alpha values ranging from 0.93 to 0.96, and an ICC value of 0.87 (95% CI = 0.70-0.99), demonstrating excellent test-retest reliability. Mean scores of individual items ranged from 0.82 ± 0.94 (Q3) to 3.23 ± 1.15 (Q11). The corrected item-total correlation ranged from 0.30 (Q2) to 0.90 (Q19). Organoleptic test scores diagnosed 41% (62 patients) with a score of 2, with increasing severity in 9% (13 patients) with a score of 5. The R-HALT exhibited robust reliability and validity in assessing the impact of halitosis among Romanian teenagers and young adults. The questionnaire is a strong tool for understanding, diagnosing, and managing halitosis in Romania, ultimately aiming to improve the quality of life of affected individuals. Further studies with diverse populations could enhance the applicability of the R-HALT.

Eugenol: In Vitro and In Ovo Assessment to Explore Cytotoxic Effects on Osteosarcoma and Oropharyngeal Cancer Cells.

Cancer is a significant health problem worldwide; consequently, new therapeutic alternatives are being investigated, including those found in the vegetable kingdom. Eugenol (Eug) has attracted attention for its therapeutic properties, especially in stomatology. The purpose of this study was to investigate the cytotoxicity of Eug, in vitro, on osteosarcoma (SAOS-2) and oropharyngeal squamous cancer (Detroit-562) cells, as well as its potential irritant effect in ovo at the level of the chorioallantoic membrane (CAM). The data obtained following a 72 h Eug treatment highlighted the reduction in cell viability up to 41% in SAOS-2 cells and up to 37% in Detroit-562 cells, respectively. The apoptotic-like effect of Eug was indicated by the changes in cell morphology and nuclear aspect; the increase in caspase-3/7, -8 and -9 activity; the elevated expression of Bax and Bad genes; and the increase in luminescence signal (indicating phosphatidylserine externalization) that preceded the increase in fluorescence signal (indicating the compromise of membrane integrity). Regarding the vascular effects, slight signs of coagulation and vascular lysis were observed, with an irritation score of 1.69 for Eug 1 mM. Based on these results, the efficiency of Eug in cancer treatment is yet to be clarified.

Assessment of the Biological Activity of Thyme Essential Oil in the Presence of the Classic Antibiotic Tetracycline.

The present study was purported to assess the activities: (i) antibacterial and synergistic against three types of both Gram-positive and Gram-negative bacteria that are susceptible to drug resistance and (ii) cytotoxic and synergistic on colorectal adenocarcinoma cells, of thyme essential oil (TEo) in combination with tetracycline (Tcyc). Chemical composition of thyme essential oil was evaluated by gas chromatography-mass spectrometry (GC-MS) method, antioxidant capacity by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay, antibacterial and synergistic properties were determined by disk diffusion method and cytotoxic activity by quantifying viable cells by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Thyme essential oil has an elevated antioxidant activity, antibacterial potential against both Gram-positive and Gram-negative bacteria, especially on S. aureus and K. pneumoniae at the highest concentration tested (50µL/mL), also having a synergistic effect when combined with tetracycline (50µL/mL TEo with 10µg/mL Tcyc). Essential oil-treated cells showed a dose-dependent reduction in colorectal adenocarcinoma cell viability, while combination with tetracycline leads to a significantly attenuated decrease in viability.

Comparative Evaluation of the Potential Antitumor of Helleborus purpurascens in Skin and Breast Cancer.

In the field of oncology, the plant kingdom has an inexhaustible supply of bioactive compounds. Phytochemical compounds isolated from Helleborus species have been found to be useful in various chronic diseases. This has brought Helleborus to the attention of medical researchers. H. purpurascens is a plant characteristic of the Carpathian area, known since ancient times for its beneficial effects. The aim of the study was to evaluate the flavonoids composition of a hydroalcoholic extract of H. purpurascens, as well as to assess its antioxidant activity and antitumor potential at the level of two healthy cell lines and four tumor cell lines. In addition, the expression of the genes involved in the apoptotic process (Bcl-2, Bad, and Bax) were evaluated. The results indicated that the extract has a high concentration of flavonoids, such as epicatechin, quercetin, and kaempferol. The extract has an increased antioxidant activity, very similar to that of the standard, ascorbic acid and cytotoxic effects predominantly in the breast cancer cell line, being free of cytotoxic effects in healthy cell lines. Underlying the cytotoxic effect is the induction of the process of apoptosis, which in the present study was highlighted by decreasing the expression of anti-apoptotic genes (Bcl-2) and increasing the expression of pro-apoptotic genes (Bad and Bax). In conclusion, the hydroalcoholic extract of H. purpurascens can be considered an important source for future medical applications in cancer therapy.

Space Maintainers Used in Pediatric Dentistry: An Insight of Their Biosecurity Profile by Applying In Vitro Methods.

Space maintainers have presented an increased interest due to their chemical composition which influences the electrochemical and electrolytic processes of the oral cavity, leading to important biological activity. The present study was purported to evaluate the biological in vitro activity of three types of space maintainers (S1, S2, and S3, differing from each other in terms of metal composition) used in pediatric dentistry, in terms of their antimicrobial effect and biosecurity profile using two types of keratinocytes (PGK: primary gingival keratinocytes, and HaCaT: human immortalized keratinocytes) by assessing the morphology, viability, cytotoxicity, and gene expression of the cells. Statistical differences were calculated by the one-way ANOVA test, followed by Tukey's post-test. Antimicrobial screening highlighted a dilution-dependent influence that, in the case of all strains tested, did not show inhibition or stimulation of bacterial growth. The in vitro evaluations revealed that the test samples did not induce important cytotoxic potential on both keratinocyte cell lines (HaCaT and PGK), with the cells manifesting no morphological alteration, a good viability rate (above 90%: PGK-S1, * p < 0.05), and a low cytotoxic activity (less than 11%: PGK, S1 *** p < 0.001 and S3 * p < 0.05; HaCaT, S1 ** p < 0.01). The data obtained in this study highlight the fact that the samples analyzed are biocompatible and do not develop the growth of the studied bacteria or encode the gene expression of primary and immortalized keratinocytes.

Vegetal Compounds as Sources of Prophylactic and Therapeutic Agents in Dentistry.

Dental pathology remains a global health problem affecting both children and adults. The most important dental diseases are dental caries and periodontal pathologies. The main cause of oral health problems is overpopulation with pathogenic bacteria and for this reason, conventional therapy can often be ineffective due to bacterial resistance or may have unpleasant side effects. For that reason, studies in the field have focused on finding new therapeutic alternatives. Special attention is paid to the plant kingdom, which offers a wide range of plants and active compounds in various pathologies. This review focused on the most used plants in the dental field, especially on active phytocompounds, both in terms of chemical structure and in terms of mechanism of action. It also approached the in vitro study of active compounds and the main types of cell lines used to elucidate the effect and mechanism of action. Thus, medicinal plants and their compounds represent a promising and interesting alternative to conventional therapy.

Phytochemical Analysis and In Vitro Cytotoxic Activity against Colorectal Adenocarcinoma Cells of Hippophae rhamnodies L., Cymbopogon citratus (D.C.) Stapf, and Ocimum basilicum L. Essential Oils.

Colorectal carcinoma (CRC) is one of the most frequently diagnosed cancer types with current deficient and aggressive treatment options, but various studied alternative therapies are able to efficiently contribute to its management. Essential oils (EOs) contain valuable compounds, with antibacterial, anti-inflammatory, and anticancer properties, which might serve as effective solutions in CRC prophylaxis or treatment. The aim of the present work was to evaluate the phytochemical composition and in vitro biological activity of essential oils derived from Hippophae rhamnoides (Hr_EO), Cymbopogon citratus (Cc_EO), and Ocimum basilicum (Ob_EO) species on HT-29 and Caco-2 human colorectal adenocarcinoma cell lines. The main compounds identified by GC-MS analysis were estragole (Hr_EO, Ob_EO), alpha- and beta-citral (Cc_EO). All tested EOs exerted a dose-dependent cytotoxicity on both cell lines by reducing the cell viability, especially in the case of Cc_EO, where at 75 µg/mL the viability percentages reached the values of 62.69% (Caco-2) and 64.09% (HT-29), respectively. The nuclear morphology evaluation highlighted significant dysmorphologies on both lines after their treatment with EOs at 75 µg/mL.

Development and Characterization of Fe3O4@Carbon Nanoparticles and Their Biological Screening Related to Oral Administration.

The current study presents the effect of naked Fe3O4@Carbon nanoparticles obtained by the combustion method on primary human gingival fibroblasts (HGFs) and primary gingival keratinocytes (PGKs)-relevant cell lines of buccal oral mucosa. In this regard, the objectives of this study were as follows: (i) development via combustion method and characterization of nanosized magnetite particles with carbon on their surface, (ii) biocompatibility assessment of the obtained magnetic nanoparticles on HGF and PGK cell lines and (iii) evaluation of possible irritative reaction of Fe3O4@Carbon nanoparticles on the highly vascularized chorioallantoic membrane of a chick embryo. Physicochemical properties of Fe3O4@Carbon nanoparticles were characterized in terms of phase composition, chemical structure, and polymorphic and molecular interactions of the chemical bonds within the nanomaterial, magnetic measurements, ultrastructure, morphology, and elemental composition. The X-ray diffraction analysis revealed the formation of magnetite as phase pure without any other secondary phases, and Raman spectroscopy exhibit that the pre-formed magnetic nanoparticles were covered with carbon film, resulting from the synthesis method employed. Scanning electron microscopy shown that nanoparticles obtained were uniformly distributed, with a nearly spherical shape with sizes at the nanometric level; iron, oxygen, and carbon were the only elements detected. While biological screening of Fe3O4@Carbon nanoparticles revealed no significant cytotoxic potential on the HGF and PGK cell lines, a slight sign of irritation was observed on a limited area on the chorioallantoic membrane of the chick embryo.

Scaffold-Type Structure Dental Ceramics with Different Compositions Evaluated through Physicochemical Characteristics and Biosecurity Profiles.

The design and development of ceramic structures based on 3D scaffolding as dental bone substitutes has become a topic of great interest in the regenerative dentistry research area. In this regard, the present study focuses on the development of two scaffold-type structures obtained from different commercial dental ceramics by employing the foam replication method. At the same time, the study underlines the physicochemical features and the biological profiles of the newly developed scaffolds, compared to two traditional Cerabone® materials used for bone augmentation, by employing both the in vitro Alamar blue proliferation test at 24, 48 and 96 h poststimulation and the in ovo chick chorioallantoic membrane (CAM) assay. The data reveal that the newly developed scaffolds express comparable results with the traditional Cerabone® augmentation masses. In terms of network porosity, the scaffolds show higher pore interconnectivity compared to Cerabone® granules, whereas regarding the biosafety profile, all ceramic samples manifest good biocompatibility on primary human gingival fibroblasts (HGFs); however only the Cerabone® samples induced proliferation of HGF cells following exposure to concentrations of 5 and 10 µg/mL. Additionally, none of the test samples induce irritative activity on the vascular developing plexus. Thus, based on the current results, the preliminary biosecurity profile of ceramic scaffolds supports the usefulness for further testing of high relevance for their possible clinical dental applications.

Midpalatal suture morphology and bone density evaluation after orthodontic expansion: a cone-bean computed tomography study in correlation with aesthetic parameters.

Maxillary expansion is one of the earliest methods of obtaining space used in the field of orthodontics. Maturing craniofacial sutures along with the increase in bone density and rigidity are main causes of high resistance of the maxilla to transversal expansion forces applied to the midpalatal suture through orthodontic appliances. Fifty-three patients, with a mean age of 16.4 years and a diagnosed transverse plane orthodontic anomaly were included in this study and divided in two groups: male group and female group. Cone-beam computed tomography (CBCT) was used for measurements conducted in order to determine bone density before and after jaw expansion in different segments of the midpalatal suture: anterior, middle and posterior. In males, slightly higher bone density values were observed in the midpalatal suture than in females before and after maxillary expansion, with average values ranging from 128.5 Hounsfield units (HU) to 672.9 HU. Bone density along the maxillary suture plays an important role in the success rate of orthodontic treatment. Assessing the palatal suture maturation on CBCT images is a very promising predictor for conventional or surgically assisted jaw expansion. Intra and extraoral pictures were used to evaluate the position of the zenith in the aesthetic area and gingival aesthetic line (GAL) class. In the study, there was a significant reduction in the number of class II, class III and class IV and an implicit increase of GAL I class that ensure a pleasant transition of the gingival level between the anterior maxillary teeth. The distribution of the gingival height in terms of the classes found prior to the orthodontic treatment remained unchanged after treatment.